Identification of proteins and genes as potential biomarkers in brown-marbled grouper (Epinephelus fuscoguttatus forsskal) resistance to Vibrio sp

The gram-negative marine bacterium, Vibrio sp. has frequently been identified as the causative pathogen responsible for the infectious disease vibriosis in the marine aquaculture industry. This disease is one of the majorc allenges facing brown-marbled grouper aquaculture, causing fish farmers globa...

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Main Author:	Low, Chen Fei
Format:	Thesis
Language:	English
Published:	2014
Subjects:	Vibrio infections Mariculture industry
Online Access:	http://psasir.upm.edu.my/id/eprint/56817/1/IB%202014%2014RR.pdf
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spelling	my-upm-ir.568172017-08-04T03:39:18Z Identification of proteins and genes as potential biomarkers in brown-marbled grouper (Epinephelus fuscoguttatus forsskal) resistance to Vibrio sp 2014-11 Low, Chen Fei The gram-negative marine bacterium, Vibrio sp. has frequently been identified as the causative pathogen responsible for the infectious disease vibriosis in the marine aquaculture industry. This disease is one of the majorc allenges facing brown-marbled grouper aquaculture, causing fish farmers globally to suffer substantial economic losses. In this study, several experiments were conducted using a range of methodologies to identify proteins and genes that are immune response-related upon Vibrio infection in brown-marbled grouper. Serum proteome profiles from two-dimensional gel electrophoresis were compared between infected grouper and control grouper after 4 hours of pathogen challenge. Differentially expressed proteins were then identified by MALDI TOF. The serum proteins that were ,ighly expressed during early Vibrio infection of grouper were putative apolipoprotein A-I, natural killer cell enhancement factor and lysozyme g.The transcriptome of brown-marbled grouper spleen was studied by RNAsequencing using Next Generation Sequencing technology. Gene expression in grouper spleen was compared between the infected grouper and control grouper. A total of 4162 unigenes were up-regulated in infected grouper, and 4988 unigenes were down-regulated. Gene ontology classification showed 338 differentially expressed genes were involved in immune system processes. Cell killing and antioxidant activity class have highest percentage of differentially expressed unigene of 34.48% and 37.74% respectively. Upregulated unigenes in the cell killing class included transporter-associated with antigen processing 2, and cytotoxic and regulatory T cell protein. KEGG pathway annotation identified eight immune-related pathways and also seven non-immune response-related pathways that were significantly enriched in differentially expressed genes. Among the most abundantly upregulated unigenes, four unigenes were found to be novel and were not annotated in any of the database. These novel unigenes warrant further identification and characterization of its function in immune response of grouper against pathogens. Lastly, brown marbled grouper fingerlings observed for seven days after experimental infection with Vibrio parahaemolyticus determined grouper susceptible to infection, with these fish having skin lesions ≥ 5mm. Grouper that were resistant to infection had no observable skin lesion. Skin lesion specimens observed under the scanning electron microscope revealed disintegration of skin around the lesion, and presence of bacterial cells under high magnification of 6000X. Serum proteome profiles were compared between the resistant and susceptible grouper by two-dimensional gel electrophoresis. Putative parvalbumin beta-2 subunit I, alpha-2-macroglobulin, nattectin and immunoglobulin light chain were identified to be differentially expressed in resistant grouper. In summary, resistance of grouper to bacterial infection involved complex mechanisms consisting of different pathways with distinct methods of activation and regulation. Genes and proteins altered in these pathways are potential markers to identify Vibrio resistant grouper as well as targets for immunomodulation and disease prevention through vaccination. It could that putative parvalbumin beta-2 subunit I, alpha-2-macroglobulin, nattectin and immunoglobulin light chain are among the important proteins participating critically in disease resistance mechanism in grouper, which are over-expressed to function collectively, thus contributing to the resistance of grouper to V. parahaemolyticus infection. Vibrio infections Mariculture industry 2014-11 Thesis http://psasir.upm.edu.my/id/eprint/56817/ http://psasir.upm.edu.my/id/eprint/56817/1/IB%202014%2014RR.pdf application/pdf en public phd doctoral Universiti Putra Malaysia Vibrio infections Mariculture industry
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English
topic	Vibrio infections Mariculture industry
spellingShingle	Vibrio infections Mariculture industry Low, Chen Fei Identification of proteins and genes as potential biomarkers in brown-marbled grouper (Epinephelus fuscoguttatus forsskal) resistance to Vibrio sp
description	The gram-negative marine bacterium, Vibrio sp. has frequently been identified as the causative pathogen responsible for the infectious disease vibriosis in the marine aquaculture industry. This disease is one of the majorc allenges facing brown-marbled grouper aquaculture, causing fish farmers globally to suffer substantial economic losses. In this study, several experiments were conducted using a range of methodologies to identify proteins and genes that are immune response-related upon Vibrio infection in brown-marbled grouper. Serum proteome profiles from two-dimensional gel electrophoresis were compared between infected grouper and control grouper after 4 hours of pathogen challenge. Differentially expressed proteins were then identified by MALDI TOF. The serum proteins that were ,ighly expressed during early Vibrio infection of grouper were putative apolipoprotein A-I, natural killer cell enhancement factor and lysozyme g.The transcriptome of brown-marbled grouper spleen was studied by RNAsequencing using Next Generation Sequencing technology. Gene expression in grouper spleen was compared between the infected grouper and control grouper. A total of 4162 unigenes were up-regulated in infected grouper, and 4988 unigenes were down-regulated. Gene ontology classification showed 338 differentially expressed genes were involved in immune system processes. Cell killing and antioxidant activity class have highest percentage of differentially expressed unigene of 34.48% and 37.74% respectively. Upregulated unigenes in the cell killing class included transporter-associated with antigen processing 2, and cytotoxic and regulatory T cell protein. KEGG pathway annotation identified eight immune-related pathways and also seven non-immune response-related pathways that were significantly enriched in differentially expressed genes. Among the most abundantly upregulated unigenes, four unigenes were found to be novel and were not annotated in any of the database. These novel unigenes warrant further identification and characterization of its function in immune response of grouper against pathogens. Lastly, brown marbled grouper fingerlings observed for seven days after experimental infection with Vibrio parahaemolyticus determined grouper susceptible to infection, with these fish having skin lesions ≥ 5mm. Grouper that were resistant to infection had no observable skin lesion. Skin lesion specimens observed under the scanning electron microscope revealed disintegration of skin around the lesion, and presence of bacterial cells under high magnification of 6000X. Serum proteome profiles were compared between the resistant and susceptible grouper by two-dimensional gel electrophoresis. Putative parvalbumin beta-2 subunit I, alpha-2-macroglobulin, nattectin and immunoglobulin light chain were identified to be differentially expressed in resistant grouper. In summary, resistance of grouper to bacterial infection involved complex mechanisms consisting of different pathways with distinct methods of activation and regulation. Genes and proteins altered in these pathways are potential markers to identify Vibrio resistant grouper as well as targets for immunomodulation and disease prevention through vaccination. It could that putative parvalbumin beta-2 subunit I, alpha-2-macroglobulin, nattectin and immunoglobulin light chain are among the important proteins participating critically in disease resistance mechanism in grouper, which are over-expressed to function collectively, thus contributing to the resistance of grouper to V. parahaemolyticus infection.
format	Thesis
qualification_name	Doctor of Philosophy (PhD.)
qualification_level	Doctorate
author	Low, Chen Fei
author_facet	Low, Chen Fei
author_sort	Low, Chen Fei
title	Identification of proteins and genes as potential biomarkers in brown-marbled grouper (Epinephelus fuscoguttatus forsskal) resistance to Vibrio sp
title_short	Identification of proteins and genes as potential biomarkers in brown-marbled grouper (Epinephelus fuscoguttatus forsskal) resistance to Vibrio sp
title_full	Identification of proteins and genes as potential biomarkers in brown-marbled grouper (Epinephelus fuscoguttatus forsskal) resistance to Vibrio sp
title_fullStr	Identification of proteins and genes as potential biomarkers in brown-marbled grouper (Epinephelus fuscoguttatus forsskal) resistance to Vibrio sp
title_full_unstemmed	Identification of proteins and genes as potential biomarkers in brown-marbled grouper (Epinephelus fuscoguttatus forsskal) resistance to Vibrio sp
title_sort	identification of proteins and genes as potential biomarkers in brown-marbled grouper (epinephelus fuscoguttatus forsskal) resistance to vibrio sp
granting_institution	Universiti Putra Malaysia
publishDate	2014
url	http://psasir.upm.edu.my/id/eprint/56817/1/IB%202014%2014RR.pdf
_version_	1747812148338229248

Identification of proteins and genes as potential biomarkers in brown-marbled grouper (Epinephelus fuscoguttatus forsskal) resistance to Vibrio sp

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